Recommended western blot primary and secondary antibody antibodies combinations:įinal Primary Antibody Concentration (µg/mL) Wash the membrane 2X for 5 minutes each with Wash Buffer.E.g 5 minutes for EveryBlot Blocking Buffer). Rinse the membrane twice with PBS and incubate with Blocking Buffer at room temperature for 1 hour with rocking (If chemical blocking buffer is used, follow manufacturer recommendation for blocking time.Note: Use a low-fluorescence PVDF membrane or nitrocellulose membrane for fluorescence imaging. If using wet transfer, use 100 V for 60 minutes.If using semi-dry transfer, use 10 V for 30 minutes.
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